壮药金樱根新颖五环三萜抗炎活性成分及效应关系的研究

The active fraction of the root of Rosa laevigata, a kind of Zhuang Nationality medicine, was confirmed to possess obvious anti-inflammatory activity in our previous study. Tentatively chemcial investigation indicated that the active fraction is rich in pentacyclictriterpene (PTT). Further more, the anti-inflammatory mechanism of the PTT obtained from RRL was found to be closely associated with inhibiting the expression of TNF-α in macrophage. On the basis of previous work, we aim to systematically investigate pentacyclictriterpene (PTT) with anti-inflammatory effect in the Radix Rosae Laevigatae. Based on the methodology integrated related method and technique of Chemistry of Traditional Chinese Medicine, Medicinal Chemistry of Serum, Computer Aided Virtual Screening of Drugs, Molecular Biology, PTT will be rapidly screened, isolated and elucidated from the active fraction of Radix Rosae Laevigatae.Meanwhile, using TNF-α as a target, molecular docking will be performed to evaluate the binding degree between the obtained compounds and the receptor with the aid of FlexX.In addition, studies on anti-inflammatory activity of the obtained compounds both in vitro and in vivo will also be carried out, then the FFL constituents with obvious anti-inflammatory activity will be resorted to large-scale preparation,and be screened using animal model of inflammation to find out some lead compounds which have both novel structures and better anti-inflammatory activities. In this way, our findings will establish solid scientific foundation, facilitating not only illustrate the therapeutic basis of this Zhuang medicine but also find new lead compounds with independent intellectual properties and development of new drugs.

课题组在前期发现壮药金樱根具有显著的抗炎活性有效部位中的五环三萜类成分可通过抑制巨噬细胞中肿瘤坏死因子（TNF-α）的表达发挥抗炎作用的基础上，本项目拟整合中药化学、药理学、分子对接、分子生物学等相关方法，对金樱根具有显著抗炎活性的三萜成分，尤其对结构新颖的五环三萜及其苷类成分进行系统的分离和结构鉴定；以TNF-a为靶点，采用FlexX程序将所得化学成分与受体进行分子对接，评价它们相互作用的强弱；同时利用细胞模型对结合较好的成分进行活性评估，应用酶联免疫法检测活性成分对细胞内TNF-α的抑制作用；对活性显著的化合物进行足量制备，采用炎症动物模型对其抗炎活性进行评估，采用免疫组化SP法检测化合物对炎症组织中TNF-α的影响；确定具有抑制TNF-α的抗炎活性成分，并深入探索此类成分的构效关系。本项目旨在阐明具有重要经济价值的金樱根抗炎药效物质基础及作用机制，为发现新型抗炎药物奠定科学基础。

大宗原料药材金樱根为蔷薇科蔷薇属植物金樱子（Rosa laevigata Michx.）、小果蔷薇（Rosa cymosa Tratt）、粉团蔷薇（Rosa multiflora Thunb. var cathayensis Rehder＆E. H. Wilson）的干燥根及根茎，具有拔毒收敛、活血化瘀、祛风驱湿等作用，是生产著名中成药三金片、金鸡胶囊的君药，也是生产妇科千金片、王老吉等著名产品的关键原料药材。项目首先重点研究了3种不同基源的金樱根药材的化学成分研究，共分离鉴定化合物96个，其中金樱子根40个，小果蔷薇根47个，粉团蔷薇9个；发现新颖五环三萜成分10个，其中金樱子根6个，小果蔷薇3个，粉团蔷薇根1个；系统开展了金樱子根、小果蔷薇、粉团蔷薇根的指纹图谱研究；开展了金樱子根、小果蔷薇根、粉团蔷薇根的质量标准研究。共发表论文11篇，其中SCI论文3篇，北大中文核心论文7篇（其中1篇为药学学报），科技核心1篇。培养硕士研究生5名，其中已经毕业4名。目前大量分离鉴定的化合物，尤其是结构新颖的五环三萜成分，为进一步继续开展计算机辅助研究、抗炎活性以及作用机理研究奠定了坚实基础，也为促进金樱根药材标准提升为药典标准，促进金樱根为原料的相关中成药三金片、金鸡胶囊、妇科千金片、王老吉的标准提升以及二次产品开发，提供了重要技术支撑。